Chikuma T, Ishii Y, Kato T, Kodama H, Hakeda Y, Kumegawa M
J Biochem. 1985 Jun;97(6):1533-9. doi: 10.1093/oxfordjournals.jbchem.a135210.
The effects of prostaglandin E2(PGE2) on the degradation of collagen and non-collagenous peptides in clonal osteoblastic MC3T3-E1 cells were investigated by using highly sensitive assay methods for PZ-peptidase, collagenase-like peptidase (CL-peptidase), dipeptidyl-aminopeptidase (DAP), leucine aminopeptidase (LAP), and post-proline cleaving enzyme (PPCE). PGE2, at concentrations of 0.1 to 4.0 micrograms/ml, doubled the PZ-peptidase and CL-peptidase activities in the cells on 24 h culturing in a dose-dependent manner. PGE2, at a concentration of 2.0 micrograms/ml, enhanced the specific activities of PZ-peptidase, CL-peptidase, DAP, LAP, and PPCE for 75 h after the start of PGE2 stimulation. The time dependent changes in PZ-peptidase and CL-peptidase activities showed similar patterns, and 3- and 2-fold increases were seen after 48 h, respectively. The protein and DNA contents gradually increased after addition of PGE2. Since the PZ-peptidase and CL-peptidase, involved in degradation of collagen peptides, were significantly induced by PGE2 in comparison with LAP and PPCE, involved in the degradation of non-collagenous peptides, these results show that PGE2 specifically stimulates induction of collagen catabolizing enzymes in clonal osteoblasts.
采用针对PZ肽酶、胶原酶样肽酶(CL肽酶)、二肽基氨基肽酶(DAP)、亮氨酸氨基肽酶(LAP)和脯氨酸后切割酶(PPCE)的高灵敏度检测方法,研究了前列腺素E2(PGE2)对克隆成骨细胞MC3T3-E1中胶原蛋白和非胶原蛋白肽降解的影响。在0.1至4.0微克/毫升的浓度下,PGE2在24小时培养中以剂量依赖的方式使细胞中的PZ肽酶和CL肽酶活性增加了一倍。在开始PGE2刺激后75小时内,浓度为2.0微克/毫升的PGE2增强了PZ肽酶、CL肽酶、DAP、LAP和PPCE的比活性。PZ肽酶和CL肽酶活性的时间依赖性变化呈现相似模式,48小时后分别增加了3倍和2倍。添加PGE2后,蛋白质和DNA含量逐渐增加。由于参与胶原蛋白肽降解的PZ肽酶和CL肽酶与参与非胶原蛋白肽降解的LAP和PPCE相比,被PGE2显著诱导,这些结果表明PGE2特异性刺激克隆成骨细胞中胶原蛋白分解酶的诱导。